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Transient expression of human and chicken progesterone receptors does not support alternative translational
P Kastner1, M T Bocquel, B Turcotte
1Laboratoire de Génétique Moléculaire des Eucaryotes, Centre National de la Recherche Scientifique Unité 184 de Biologie Moléculaire et de Génie Génétique, Strasbourg, France.
Abstract:
Two isoforms (A and B) of the human (hPR) and chicken (cPR) progesterone receptors originate from a single PR gene. cPR form A results from initiation of translation at a downstream ATG codon (ATG2) which in the cPR cDNA-deduced open reading frame is found 128 amino acids C-terminal to and in-frame with the first ATG codon (ATG1) that gives rise to form B. Our recent observation of an abundant cPR mRNA which encodes only form A suggested to us that the two isoforms are translated from different transcripts (Jeltsch, J. M., Turcotte, B., Garnier, J. M., Lerouge, T., Krozowski, Z., Gronemeyer, H., and Chambon, P. (1990) J. Biol. Chem. 265, 3961-3974). This view is, however, at variance with data obtained by transient transfection with expression vectors containing most of the cPR cDNA (downstream of nucleotide +53) since both isoforms were generated in transiently transfected COS cells (Conneely, O. M., Kettelberger, D. M., Tsai, J. J., Schrader, W. T., and O'Malley, B. W. (1989) J. Biol. Chem. 264, 14062-14064). To further support our above conclusion, vectors containing either hPR or cPR cDNAs were introduced into HeLa and COS-1 cells. Only hPR form B originated from a vector containing the entire cDNA (containing nucleotides 1 to approximately 4400), and form A was produced only from a vector expressing hPR transcripts (nucleotides 814 to approximately 4400) lacking ATG1. Vectors expressing the 5'-untranslated and coding region of the cPR mRNA (nucleotides 29-2921) generated only traces of form A in the two cell lines. Similar traces of form A were observed in COS-1 cells transfected with a vector lacking the 5'-untranslated region. Collectively, these results do not support the hypothesis that similar amounts of the two PR isoforms are generated by alternative initiation of translation on a single PR transcript. We discuss data indicating that for hPR and cPR, isoforms A and B are in fact translated from different mRNAs.